Qian Li1, Ping Huang2, Weimin Chen3, Jiarui Bi4. 1. Department of Radiology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, No. 1095 Jiefang Road, Wuhan, 430030, Hubei, China. 2. Department of Stomatology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, No. 1095 Jiefang Road, Wuhan, 430030, Hubei, China. huangping_hust@hotmail.com. 3. Department of Stomatology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, No. 1095 Jiefang Road, Wuhan, 430030, Hubei, China. 4. Division of Periodontology, Diagnostic Sciences, and Dental Hygiene, Herman Ostrow School of Dentistry, University of Southern California, Los Angeles, CA, USA.
Abstract
OBJECTIVE: The aim of this study was to investigate the molecular mechanism of human bone marrow mesenchymal stem cells (hMSCs) secreting miR-26a exosomes on the function of skin fibroblasts. METHODS: Exosomes from hMSCs were extracted and identified by transmission electron microscopy, particle size was analyzed and protein markers were detected. Then, the exosomes were co-cultured with human skin fibroblasts (BJ). CCK-8, Annexin V/P and Transwell assays were used to detect the proliferation, apoptosis, and migration of BJ cells. In addition, the expressions of miR-26a, related proteins, and related inflammatory factors were detected by qRT-PCR, western blotting, and ELISA. RESULTS: Compared with the high glucose group, the proliferation rate, migration rate, and the expression of α-SMA, bcl-2, TLR4, NF-κB, TNF-α, IL-6, IL- and IL-1 were significantly decreased in the high glucose + MSC-Exo-miR-26a mimics group, while the apoptosis rate and the expression of miR-26a, cleaved-caspase 3, cleaved-caspase 9 and Bax were significantly increased. The results of the high glucose + MSC-Exo-miR-26a inhibitor group were the opposite. CONCLUSION: These results suggest that hMSCs cells secreting miR-26a exosomes inhibited the proliferation, migration, and transdifferentiation of high glucose-induced BJ cells, and promoted cell apoptosis, which may be related to the TLR4/NF-κB signaling pathway.
OBJECTIVE: The aim of this study was to investigate the molecular mechanism of human bone marrow mesenchymal stem cells (hMSCs) secreting miR-26a exosomes on the function of skin fibroblasts. METHODS: Exosomes from hMSCs were extracted and identified by transmission electron microscopy, particle size was analyzed and protein markers were detected. Then, the exosomes were co-cultured with human skin fibroblasts (BJ). CCK-8, Annexin V/P and Transwell assays were used to detect the proliferation, apoptosis, and migration of BJ cells. In addition, the expressions of miR-26a, related proteins, and related inflammatory factors were detected by qRT-PCR, western blotting, and ELISA. RESULTS: Compared with the high glucose group, the proliferation rate, migration rate, and the expression of α-SMA, bcl-2, TLR4, NF-κB, TNF-α, IL-6, IL- and IL-1 were significantly decreased in the high glucose + MSC-Exo-miR-26a mimics group, while the apoptosis rate and the expression of miR-26a, cleaved-caspase 3, cleaved-caspase 9 and Bax were significantly increased. The results of the high glucose + MSC-Exo-miR-26a inhibitor group were the opposite. CONCLUSION: These results suggest that hMSCs cells secreting miR-26a exosomes inhibited the proliferation, migration, and transdifferentiation of high glucose-induced BJ cells, and promoted cell apoptosis, which may be related to the TLR4/NF-κB signaling pathway.